Fig 1: Met7 contributes to the stability of the genome. Inactivation of Met7 leads to folate depletion causing limiting dTTP and dGTP and accumulation of dUTP. A high dUTP/dTTP ratio promotes uracil incorporation into DNA. Uracil is removed from DNA by Ung1, however the high dUTP/dTTP ratio leads to futile uracil excision/incorporation cycles resulting in abasic sites and DSBs. Limiting dNTPs (dTTP and dGTP) may also affect DNA repair capacity and increase the frequency of mutations.
Fig 2: Inactivation of Met7 results in a high dUTP/dTTP ratio that causes increased uracil incorporation into DNA. HPLC quantification of NTP (A) and dNTP (B) concentrations in cell extracts of indicated strains. Numbers on top in green or red, indicate the fold increase or decrease, respectively, relative to WT levels. In (B), cell extracts were (+) or not (−) treated with recombinant human Dut1 prior quantification of dNTPs. ‘nd’ (or ‘not detectable’) indicates dUTP concentrations below our detection limit (≤3 pmol dUTP). (C) Schematic diagram illustrating approach used in (D) for the detection of incorporated uracil into genomic DNA. (D) Genomic DNAs isolated from the indicated strains were (or not) digested with UDG + Ape1 enzymes and subsequently visualized by agarose gel electrophoresis.
Supplier Page from Abcam for Recombinant Human dUTPase protein (BSA and azide free)